Methods in Molecular Biology Volume 958, 2013, pp 211-222

Agrobacterium-Mediated Transformation of Cotton (Gossypium hirsutum) Shoot Apex with a Fungal Phytase Gene Improves Phosphorus Acquisition

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Abstract

Cotton is an important world economic crop plant. It is considered that cotton is recalcitrant to in vitro proliferation. Somatic embryogenesis and plant regeneration has been successful by using hypocotyl, whereas it is highly genotype dependent. Here, a genotype-independent cotton regeneration protocol from shoot apices is presented. Shoot apices from 3- to 5-day-old seedlings of cotton are infected with an Agrobacterium strain, EHA105, carrying the binary vector pC-KSA contained phytase gene (phyA) and the marker gene neomycin phosphotransferase (NPTII), and directly regenerated as shoots in vitro. Rooted shoots can be obtained within 6–8 weeks. Plants that survived by leaf painting kanamycin (kan) were ­further analyzed by DNA and RNA blottings. The transgenic plants with increased the phosphorus (P) acquisition efficiency were obtained following the transformation method.